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- 저자 : Huiwen Yan (검색결과 2 건)
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Huiwen Yan,Lei Sun,Jinge Huang,Yixing Qiu,Fuchao Xu,Riming Yan,Du Zhu,Wei Wang,Jixun Zhan 한국미생물학회 2018 The journal of microbiology Vol.56 No.11
A new type III polyketide synthase gene (Ssars) was discovered from the genome of Shiraia sp. Slf14, an endophytic fungal strain from Huperzia serrata. The intron-free gene was cloned from the cDNA and ligated to two expression vectors pET28a and YEpADH2p-URA3 for expression in Escherichia coli BL21(DE3) and Saccharomyces cerevisiae BJ5464, respectively. SsARS was efficiently expressed in E. coli BL21(DE3), leading to the synthesis of a series of polyketide products. Six major products were isolated from the engineered E. coli and characterized as 1,3-dihydroxyphenyl- 5-undecane, 1,3-dihydroxyphenyl-5-cis-6 -tridecene,1,3-dihydroxyphenyl- 5-tridecane, 1,3-dihydroxyphenyl-5-cis-8 - pentadecene, 1,3-dihydroxyphenyl-5-pentadecane, and 1,3- dihydroxyphenyl-5-cis-10 -heptadecene, respectively, based on the spectral data and biosynthetic origin. Expression of SsARS in the yeast also led to the synthesis of the same polyketide products, indicating that this enzyme can be reconstituted in both heterologous hosts. Supplementation of soybean oil into the culture of E. coli BL21(DE3)/SsARS increased the production titers of 1–6 and led to the synthesis of an additional product, which was identified as 5-(8 Z,11 Z-heptadecadienyl) resorcinol. This work thus allowed the identification of SsARS as a 5-alk(en)ylresorcinol synthase with flexible substrate specificity toward endogenous and exogenous fatty acids. Desired resorcinol derivatives may be synthesized by supplying corresponding fatty acids into the culture medium.
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Comparison of leaf transcriptomes of cassava “Xinxuan 048” diploid and autotetraploid plants
Ling Yin,Junjie Qu,Huiwen Zhou,Xiaohong Shang,Hui Fang,Jiang Lu,Huabing Yan 한국유전학회 2018 Genes & Genomics Vol.40 No.9
Polyploidy breeding of cassava has been used to improve cassava traits over the past years. We previously reported in vitro induction of tetraploids in the cassava variety “Xinxuan 048” using colchicine. Significant differences in morphology and anatomy were found between the diploid and tetraploid plants. However, very little is known about the transcriptome difference between them. In this study, morphological and physiological characteristics including leaf thickness, plant height, internode length, chlorophyll content, and photosynthetic capacity were measured. Further, we investigated and validated the difference in gene expression patterns between cassava “Xinxuan 048” tetraploid genotype and its diploid plants using RNA sequencing (RNAseq) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Significant differences in morphology and physiology were observed during tetraploidization. A comparison revealed that tetraploidy induced very limited changes in the leaf transcriptomes of cassava “Xinxuan 048” diploid and autotetraploid plants. However, the differentially expressed genes (DEGs) between 2× and 4× plants, especially those upregulated in 4× plants, were strongly associated with hormonal and stress responses. Large changes in morphology and physiology between the diploid cassava “Xinxuan 048” and its autotetraploid were not associated with large changes in their leaf transcriptomes. Moreover, the differently expressed genes related to the regulation of gibberellin and brassinosteroids potentially explained why the plant height and internode length of 4× plants became shorter. Collectively, our results suggest that 4× cassava is potentially valuable for breeding strains with improved stress resistance.
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